Human NEK6 knockout HeLa cell lysate
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NEK6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3 and 304 bp deletion in exon3 and Insertion of the selection cassette in exon3.
View Alternative Names
Highly similar to cell cycle protein kinase CDC5/MSD2, NEK6_HUMAN, NIMA (Never In Mitosis Gene A) Related Kinase 6, Never in mitosis A-related kinase 6, NimA-related protein kinase 6, Protein kinase SID6-1512, Putative serine threonine protein kinase, SID6 1512, Serine/threonine-protein kinase Nek6
- WB
Lab
Western blot - Human NEK6 knockout HeLa cell lysate (AB258072)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : NEK6 knockout HeLa cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lanes 1-3 : Merged signal (red and green). Green - ab109177 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.
ab109177 Anti-NEK6 antibody [EPR5282] was shown to specifically react with NEK6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265910 (knockout cell lysate ab258072) was used. Wild-type and NEK6 knockout samples were subjected to SDS-PAGE. ab109177 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Lanes 1 - 3:
Anti-VEGF Receptor 3 antibody (<a href='/en-us/products/unavailable/vegf-receptor-3-antibody-ab10977'>ab10977</a>) at 1/1000 dilution
Lanes 1 - 3:
Western blot - Anti-NEK6 antibody [EPR5282] (<a href='/en-us/products/primary-antibodies/nek6-antibody-epr5282-ab109177'>ab109177</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
NEK6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human NEK6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nek6-knockout-hela-cell-line-ab265910'>ab265910</a>)
Lane 3:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 15 kDa,36 kDa,54 kDa,66 kDa,88 kDa,92 kDa
Observed band size: 15 kDa,36 kDa,56 kDa,88 kDa,92 kDa
false
- WB
Lab
Western blot - Human NEK6 knockout HeLa cell lysate (AB258072)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : NEK6 knockout HeLa cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lanes 1-3 : Merged signal (red and green). Green - ab133494 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.
ab133494 Anti-NEK6 antibody [EPR5283] was shown to specifically react with NEK6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265910 (knockout cell lysate ab258072) was used. Wild-type and NEK6 knockout samples were subjected to SDS-PAGE. ab133494 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-NEK6 antibody [EPR5283] (<a href='/en-us/products/primary-antibodies/nek6-antibody-epr5283-ab133494'>ab133494</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
NEK6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human NEK6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nek6-knockout-hela-cell-line-ab265910'>ab265910</a>)
Lane 3:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 36 kDa,53 kDa
Observed band size: 36 kDa,53 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human NEK6 knockout HeLa cell lysate (AB258072)
Allele-2 : 1 bp insertion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human NEK6 knockout HeLa cell lysate (AB258072)
Allele-1 : 304 bp deletion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human NEK6 knockout HeLa cell lysate (AB258072)
Allele-3 : Insertion of the selection cassette in exon3
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NEK6 plays a critical role in mitotic cell division by promoting progression through the G2/M phase of the cell cycle. Although NEK6 does not form part of a classical protein complex it does interact with various substrates to ensure successful mitosis. It stabilizes microtubule organization ensuring proper chromosomal segregation and cell cycle progression by facilitating microtubule nucleation and stability.
Pathways
NEK6 contributes to cell cycle control and microtubule dynamics being important within the context of the DNA damage response and repair pathways. NEK6 is associated with the p53 signaling pathway where it functions alongside related proteins such as NEK7. These kinases coordinate cellular responses to stress and damage ultimately maintaining genomic stability and preventing aberrant cell division.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-NEK6 antibody [EPR5283] (AB133494)](https://content.abcam.com/products/images/nek6-antibody-epr5283-ab133494--western-blot-img104373.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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